Life Sciences Contribution 9 8 Royal Ontario Museum Electrophoretic Patterns of Serum Proteins of Neotropical Bats (Chiroptera) Dario Valdivieso and J. R. Tamsitt ROM Se = - as - - zs ——j eee lead ~ 3 nd . ——= : ~ — tuscan — a . = = : “ = " - 7 ~< 4 - Sa c = a - = 2 = “ 5 7 — — es - = - = x ~- \ Ad SPARE vie | mes Wy if Pabets FAD eit hil : té We ik if “ — et - - ——— —— <7. ——— ~ ~ 0 Ae = - - ———— a a tea Pe ga —— — — a ————— ~_ - ——— eee = a a a a neal = ey ig ————— _ — i 7 - = <~ — = ee -_ -_ _ _ ~ - Peet ew i 2 _ — —— —— _ a - - 2 - r a er PL ee ge all [— _ ” _ i Rum = oe ———-— rg ee ne ae a 0 ee = - : = = - “ = = _ < — a Sp —_ = 6 a (Ge s| t'8 a ve ee = aie! SIE veer ONL a O'S ye v € Cc *(soumatyxes “qs = X) sulajo1d windas JO suoneUus.U0D sATILIOY OC (ONY GC 8°C TOL L'v9 VCS C59 9°79 vos 80S v9S COS O'CS Cc 8V c 6Pr 00S 86S urwungTy (88°0 ‘88°0) 88°0 68 °0 (68°0 ‘68°0) 68'0 (LL°0-0L'0) cO'0 = ¥L0 (78'0-18'0) 10°0 + ¢8°0 S80 ulunqye jo AyI]Iqouw DATILIOY 5 ynpe I wuni}i] 2 inpe I DAIUANIS 1UDULPIA 35 ynpe 9 snjjkydouopw & plavis € DUIDIAOS & 1npe I vévydosso]y & Plavis t © snjay - 2 yupe sunoAd ¢ 4O]OISIp 2 ynpe bp snwojsoj] VL 8's $°89 7O';O + 760 PP Wnpe (4 Cc SNSSOJOW Results ELECTROPHEROGRAMS OF SERA Four to eight fractions were visible in electrophoretic patterns of chirop- teran serum proteins (Figs. 1 and 2), and patterns of 18 species are sum- marized in Fig. 3. Quantitative aspects of profiles are compared in Table I. When polymorphism occurred, it involved fractions in the alpha and beta globulins. Interspecific differences were observed in the total number of fractions and in their relative mobilities (Fig. 3). Although varying in electrophoretic mobility among species, albumin, the most intense and rapidly migrating zone, and the gamma globulin fraction were mono- morphic; consequently, although included in the total number of fractions, they are not discussed here. Intraspecific variation in the number of fractions in the alpha and beta globulin regions occurred in seven species (Table 1; Fig. 3). In Phyllo- stomus hastatus (Pallas) sera separated into five or six fractions (Fig. 1B,c). In the alpha region two fractions were present in a male and a female, whereas in another male and female three fractions were present. Five fractions, including two in the alpha and one in the beta region, were seen in adult P. discolor (Wagner). Serum of a male fetus, however, showed four fractions, with only one in the alpha region (Fig. 1D). Four or five fractions were present in Glossophaga soricina (Pallas). An adult, non- parous female possessed a single alpha globulin and a single beta globulin, whereas in a gravid female an additional alpha fraction was present (Fig. icy. In Artibeus jamaicensis Leach serum protein fractions varied from five to eight (Figs. 1A,E; 2A). Adult males had the greatest number (eight), with four fractions in the alpha region and two in the beta globulin region. In two adult females fractions numbered seven (three alpha and two beta). A lactating female had only five fractions, with two alphas and a single beta, a pattern similar to that of three young adult males in which the third and fourth alpha fractions were absent. Of the age groups, young adult males were the most variable; two had seven fractions (three alpha and two beta), one had six (two alpha and two beta), and three had five (two alpha and one beta fractions). One young adult female had six fractions, two alpha and two beta globulins that were comparable to those of one of the young adult males. Five or six fractions were found in sera of Artibeus cf. jamaicensis (Fig. 2B). Six fractions were found in an adult male and a gravid female, with two in the alpha region and two in the beta region. Only five fractions were seen in a lactating female, whose electropherogram was identical to that of other adults except for the absence of one of the beta globulins. Serum fractions of Artibeus lituratus Olfers from Colombia and Vene- zuela varied in number from four to six (Figs. 1c; 2B,c). Adult males possessed five or six fractions; those with five fractions had either two fractions in the alpha region and one in the beta region (one male from Colombia, one male from Venezuela) or one fraction in the alpha region 8 and two fractions in the beta region (one male from Colombia). Males with six fractions possessed two in the alpha and two in the beta regions (four males from Colombia, five from Venezuela). Serum fractions in females varied from four to six. One adult female (Venezuela) had only four fractions, including a single alpha and a single beta globulin. One adult, non-parous female (Venezuela) had six fractions, with two in the alpha and two in the beta regions. One gravid female (Colombia) and one adult, non-parous female (Venezuela) possessed five fractions, including one in the alpha and two in the beta regions. Samples from Colombia and Venezuela showed the same degree of variability, and numbers of serum fractions varied in individuals of each of the disjunct populations. In Carollia perspicillata Linnaeus five or six serum fractions were present (Fig. 1c,F). Patterns in two males and two gravid females were identical, consisting of three fractions in the alpha and one in the beta region. In one gravid female, however, only two fractions were present in the alpha region. Comparison of percentage composition of serum proteins by fraction according to sex and age without specific allocation resulted in only one statistically significant difference. The percentage (13.8) of fractions in the alpha region was significantly less in 14 adult, non-parous females than that (29.8) in 13 gravid females (P <= 0.025), although no other differ- ences were found. Nor were significant differences (P < 0.05) found be- tween adult and young adult males, between adult males and adult females, between adult and lactating females, or between adult and young adult females. RELATIVE MOBILITY OF ALBUMIN Among the species studied, none of the albumins migrated anodal to the NHS control (Table 1; Figs. 1-3). Artibeus cf. jamaicensis, A. lituratus, A. phaeotis (Miller), Stenoderma rufum St. Hilaire, Carollia perspicillata, and Desmodus rotundus (Geoffroy) possessed albumins that had the same mobility as the NHS control. Glossophaga soricina, Monophyllus redmani Leach, Sturnira lilium (Geoffroy), Brachphylla cavernarum Gray, Arti- beus jamaicensis, Erophylla bombifrons (Miller) (Phyllostomatidae), Eptesicus fuscus Palisot de Beauvois (Vespertilionidae), Molossus molossus (Pallas), and M. fortis Miller (Molossidae) possessed albumins with simi- lar mobilities that were cathodic to the NHS control. Phyllostomus discolor and P. hastatus possessed albumin mobilities that differed from other species by being the most cathodic (Fig. 3). Likewise, the mobility of the albumin of P. discolor was more cathodic than that of P. hastatus. The mobility of the albumin of Pteronotus parnellii (Gray) (Mormoopidae) was slightly cathodic to albumins of the species of Phyllostomus studied. PROTEIN CONCENTRATION Values of total protein content (g/100 ml) of sera of adult bats ranged from 4.08 in Monophyllus redmani to 6.55 in Phyllostomus discolor and Artibeus jamaicensis, but most values were between 5 and 6 (Table 11). 9 Fig. 1 10 gy TS: at 0 Smears 26 1 Ch RA 5. 6 Electrophoretic patterns of serum proteins of bats. The arrow indicates the point of application. A. 1, Pteronotus parnellii @; 2, Artibeus jamaicensis 9; 3, Stenoderma rufum 9; 4, Erophylla bombifrons 9; 5,7, normal human serum (NHS); 6, Eptesicus fuscus 9; 8, Molossus fortis 9. All from Puerto Rico. B. Phyllostomus hastatus (Venezuela): 1, 6, NHS; 2, 3, 29; 4, 5, 264. c. 1, Phyllostomus hastatus 2 (Venezuela); 2, Glossophaga soricina 9 (Colombia); 3, Carollia perspicillata 6 (Colombia); 4, Sturnira lilium 6 (Colombia); 5, Artibeus lituratus 6 (Colombia); 6, Desmodus rotundus @ (Colombia); 7, Molossus molossus 6 (Colombia); 8, NHS. D. Phyllostomus discolor (Colombia): 1, 6, NHS: 2, fetus ¢; 3, young adults3 + 4,-6:5 55.2: Differences in our samples unequivocally attributable to age were negligible except in P. discolor, in which values for an adult were greater than those of a fetus. Differences in protein concentration in young adult and adult A. jamaicensis were insignificant (P = 0.05), as were these values for adult males and females. The relative proportions of protein (g%) of the serum fractions were variable and apparently influenced to some extent by age or reproductive condition. Adult and young adult A. jamaicensis did not differ significantly (0.10 < P > 0.05) in percentages of beta and gamma globulins, nor did other protein fractions differ significantly between age groups or between sexes. In P. discolor differences between the adult and fetus were primarily in the reduced percentages of albumin and alpha fractions of the latter, although per cent protein of the gamma of adult and fetus were compar- able. Young adult and adult A. jamaicensis were only near-significantly different (P < 0.05) in per cent protein of the gamma fraction. The great- est percentage of protein in the gamma region was found in a lactating Brachyphylla cavernarum and the least in a young adult male A. jamaicensis, an adult male Desmodus rotundus, and an adult male Molossus fortis (Table 11). Percentages of protein in the alpha and beta fractions varied remarkably and could not be correlated with age, sex, or taxon. Discussion Species similarities established by serum protein electropherograms (Fig. 3) do not agree with existing taxonomic arrangements. Bats of the family Phyllostomatidae have a particularly wide range of variation in serum protein patterns similar to that reported for karyotypes by Baker (1970). Although no albumin polymorphism was observed in populations of species studied, differences and similarities were noted among several groups of species (Table 1; Fig. 3). None of the albumins studied migrated anod- ically to the NHS control; in all species albumin had the same mobility as the control or was cathodic to it. According to mobility of albumin, four groups could be distinguished: (1) Pteronotus parnellii (Mormoopidae) ; (2) the Phyllostomatinae represented by Phyllostomus hastatus and P. discolor, which, although differing in mobilities, had the most cathodic albumins of the bats studied; (3) a group of phyllostomatid bats whose albumin mobilities were identical with NHS albumin: three species of Artibeus (A. cf. jamaicensis, A. lituratus, and A. phaeotis), the single species of Stenoderma, one species of Carollia, and the vampire bat Des- modus rotundus; and (4) a larger group of bats representing three families whose albumins were slightly cathodic to that of the control and similar to that of P. parnellii: the glossophagine bats Glossophaga soricina and Monophyllus redmani, the stenodermine bats Sturnira lilium, A. jamaicen- E. 1, 2, Artibeus jamaicensis (1, 9; 2, 6); 3, NHS; 4, Molossus fortis 3; 5, Monophyllus redmani 9°; 6, Brachyphylla cavernarum 9. All from Puerto Rico. F. Carollia perspicillata (Colombia): 1, NHS; 2-4, gravid 99; 5, ¢. Il e4 ~~. &2 to ; /) GC id ‘ a Pa = e2ep* ,g © anee Ss CSP ee GP eee ar exe Balh ny nat Pee nen Te wee eee a ee ES oe (5 Sete CA AHMED Aeead OME Gecegp ES wet OE amt ane Comb Hes A ag Sib As GAG Tels. 02.7 {geo die hod ae ete Fig. 2 Electrophoretic patterns of serum proteins of bats. The arrow indicates the 12 point of application. A. Artibeus jamaicensis (Puerto Rico): 1, 2, 9 9; 3, 4, 6-8, ¢ 4; 5, normal human serum (NHS). B. 1, NHS; 2, 3, Artibeus lituratus 6 2; 4, Artibeus lituratus gravid 9; 5, 7, Artibeus cf. jamaicensis lactating 9; 6, Artibeus cf. jamaicensis gravid 2; 8, Artibeus phaeotis lactating 9. All from Colombia. c. 1, 8, NHS; 2-4, Artibeus lituratus ¢ 6 (Colombia); 5-7, A. lituratus 3 3 (Venezuela). Dp. 1, 5, NHS; 2, 3, 6-8, Erophylla bombifrons (2, 2; 3, 6-8, 24); 4, Pteronotus parnellii 2. All from Puerto Rico. E. Desmodus rotundus (Colombia): 1, NHS; 2, 3, 99; 4, 5, gravid 2 9; 6;.7.,30°6% F. Molossus molossus (Colombia): 1, 6, NHS; 2, 3, 66; 4, 9; 5, lactating 9. sis, and Brachyphylla cavernarum, the phyllonycterine bat Erophylla bom- bifrons (Phyllostomatidae), Eptesicus fuscus (Vespertilionidae), and Molossus molossus and M. fortis (Molossidae). It is noteworthy that albumin mobilities of the two congeneric species Phyllostomus hastatus and P. discolor were significantly different (0.005 < P > 0.001). Likewise, although mobilities of albumin of the three species of mainland Artibeus were similar or not significantly different (P > 0.05), mobilities of albumin of A. jamaicensis from Puerto Rico and of mainland A. lituratus and A. cf. jamaicensis differed significantly (0.01 ene 0).025):, The status of bats previously placed in the subfamily Chilonycterinae has been questioned (Koopman and Jones, 1970; Jones and Genoways, 1970), and species of the genera Pteronotus and Mormoops were recently placed by Smith (1972) in a distinct family, the Mormoopidae. Although P. parnellii was the only taxon of this family we studied, the mobility of the albumin of this species from Puerto Rico was intermediate between the phyllostomatid bats of the subfamily Phyllostomatinae (P. hastatus and P. discolor) on the one hand and the phyllostomatid bats of the sub- families Stenoderminae (G. soricina, M. redmani, S. lilium, B. cavernarum, A. jamaicensis) and Phyllonycterinae (FE. bombifrons), and of the families Vespertilionidae (E. fuscus) and Molossidae (M. molossus and M. fortis) on the other. G. soricina, M. redmani, and S. lilium have fewer globulin fractions than P. parnellii, whereas B. cavernarum and A. jamaicensis have a greater number. Hemoglobins (Valdivieso et al., 1969) and immunologic properties of serum proteins (Gerber and Leone, 1971) of species of Pteronotus differ from those of bats of the Phyllostomatidae. But the serum protein pattern of P. parnellii does not differ markedly from other bats studied and consequently does not support the contention of Smith (1972) that this group represents a distinct family. The two species of the subfamily Phyllostomatinae, Phyllostomus has- tatus and P. discolor, have the most cathodic albumin of all bats studied and differ from other taxa in the Phyllostomatidae in this character. In numbers and mobilities of protein fractions, P. hastatus is more similar to Pteronotus parnellii of the Mormoopidae than to either P. discolor of the same genus or to other taxa of the Phyllostomatidae. Adult P. discolor have one less fraction than P. hastatus and are unique among the bats studied. These two species differ in a number of morphological characters and food habits (Walker, 1968) and as well differ in susceptibility to dental disease (Phillips and Jones, 1970) and in thermoregulatory abilities (McNab, 1969). Likewise, although chromosomes of the two taxa are similar, they differ in one pair of autosomes, an exception to the undifferentiated karyo- types typically encountered among different species of the same genus (Kib- lisky, 1969). Electrophoretic patterns of hemoglobins, however, do not differ between the two species or among bats of other subfamilies of the Phyllostomatidae (Tamsitt and Valdivieso, 1969). Sturnira lilium, in albumin mobility, does not differ from Brachyphylla cavernarum, Artibeus jamaicensis, Monophyllus redmani, and Erophylla I3 SUOTJOLIJ OM} IO UO, x SUOT}OVIJ OM}, oe ———— LO'T 6r°0 £9°0 ¥e0 c8e sc9 a’ I (SHN) wnisg ueuMy [euWION evo hs || «VSO Lv'0 EOE Ly'9 ? ynpe I SIj40f snssojow 80 «CSO LL‘0 $c0 V2 % L's Y 3yhpe I snpunjod snpowsaqd 180 LO'T «CC O «810 66°€ LE9 ? ynpe I suodfiquiog vj]jXydosy (280 ‘L9°0) (L460 ‘L8°0) (8€°0 ‘€€'0) CE0r6c 0) S(CO'C 860) “(Secs ces) LL‘0 «C60 9¢°0 +e 00°€ ges d Wupe Cc (78'0-9€'0) (Sr I-S0'T) (L8°0-T€'0) Che 0-21,0) (OL'€-89°7) (€7°9-61'S) LOO = 790 «*10°0 = 77L «x000 + 690 «£00 + ETO CFO + SIE C7O + EBS ? ynpe sunoA 9 (00°I-r2'0) Coir 10'1) (79'°0-€h'0) (LE0-47'0) (Tr €-79'7) (SS°9-61'°S) 80°0 + 160 «00 + LOT «90'0 + 9S°0 epOii==1Ce0 “SCO woe LV 2S P Wupe € sisuaawuvl snaquapy Ly «SSO 08°0 *Cv'0 60°€ €0°9 & Suyeyory if wunivusaavd vjpkyddyovig 6L'0 89°0 810 6c 0 Alar 80'P 6 yupe I 1upuipad snjjkydouopw 96°0 66°0 09°0 =~ y9'C CLs P Snyez I c6'0 €8°0 90 Lv'0 OLE ¢s°9 2 yynpe I 4OJOISIp snwojso]]Kyd St te Si SE ee a a a a ee ee Se es Oe ee ee re ee ee ee ee A J “20 ee ulin] y (jw QOI/3) xes pur ody ‘ON sa1oeds ulgjOo1d [e}0} %3 ulgj01d [vJO], *(sowelj}x9 “AS + X) Bios Jeq JO suONseIZ Jo suoljiodoid sajelor pue ulojoid [ejOy, “TT AGeL 14 bombifrons. In the number of fractions in the globulin regions, however, S. lilium is more similar to Stenoderma rufum and Monophyllus redmani than to other phyllostomatid bats. Although once considered to be a separ- ate subfamily, the Sturnirinae (Miller, 1907), Koopman and Jones (1970) placed bats of the genus Sturnira in the Sternoderminae but in a separate tribe, the Sturnirini. In karyotypes (Gardner and O’Neill, 1969), electro- phoretic properties of hemoglobins (Tamsitt and Valdivieso, 1969), and immunological properties of sera (Gerber and Leone, 1971), species of Sturnira have strong affinities with bats of the subfamily Stenoderminae. The serum protein pattern of S. lilium closely resembles that of Carollia perspicillata (Fig. 3), differing primarily in albumin mobility. Our data thus indicate a relationship with the carolline and glossophagine bats as well as with stenodermine bats. Species of the genus Brachyphylla were recently placed as a tribe of the subfamily Stenoderminae (Phyllostomatidae) by Koopman and Jones (1970) but as a member of the Phyllonycterinae by Silva and Pine (1969). In albumin mobility B. cavernarum is similar to phyllostomatids from main- land South America (G. soricina and S. lilium) and from Puerto Rico (M. redmani, A. jamaicensis, and E. bombifrons). In globulin fractions, however, B. cavernarum differs primarily from the above in number and position except in those individuals of A. jamaicensis that possess six fractions. Although species of the genera Erophylla and Brachyphylla are similar in morphology and behaviour (Silva and Pine, 1969), karyotypes (Baker and Lopez, 1970), hair structure (Benedict, 1957), and possess in common host-specific spinturnicid mites (Dusbabek, 1969), the serum protein pattern of B. cavernarum is surprisingly unlike that of E. bombi- frons and more similar to that of the vespertilionid bat Eptesicus fuscus. Erophylla bombifrons differs from other phyllostomatids studied in the same way that B. cavernarum differs. These two species are similar in albu- min mobility but differ in mobilities and numbers of globulin fractions. Lactate dehydrogenase isoenzymes of B. bombifrons differ from those of Puerto Rican A. jamaicensis (Valdivieso et al., 1968), and although species of Erophylla unquestionably are taxonomically distinct from stenodermine bats, results from electrophoretic studies of serum proteins do not confirm the conclusion of Silva and Pine (1969). Of the four species of Artibeus, the insular A. jamaicensis differs from the mainland A. lituratus, A. cf. jamaicensis, and A. phaeotis by having a slower-moving albumin and by a greater number of protein fractions (eight as opposed to 4-6). A. lituratus and A. cf. jamaicensis are similar in mobili- ties of protein fractions, but although the slower fraction in the beta region may or may not be present in individuals of both species, only in A. cf. jamaicensis is the fastest fraction in the alpha region always present. A. phaeotis, although having the same albumin mobility as A. lituratus and A. cf. jamaicensis, differs from these two taxa by the absence of the di- morphic, slower beta fraction and by the absence of the faster fraction in the alpha region. / 12) NORMAL HUMAN SERUM FAMILY MORMOOPIDAE Pteronotus parnellii FAMILY PHYLLOSTOMATIDAE Phyllostomus hastatus Phyllostomus discolor Glossophaga soricina Monophyllus redmani Sturnira lilium Brachyphylla cavernarum Artibeus jamaicensis Artibeus cf. jamaicensis lituratus Artibeus Artibeus phaeotis Stenoderma rufum Carollia perspicillata Erophylla bombifrons rotundus Desmodus FAMILY VESPERTILIONIDAE Eptesicus fuscus FAMILY MOLOSSIDAE Molossus molossus Molossus fortis NORMAL HUMAN SERUM Comparison among these congeneric species presents striking differences that raise questions of functional necessity and diversity of serum proteins. In A. phaeotis, the limited number of fractions contrasts with the marked complexities of fractions of sera in all individuals of A. lituratus and A. jamaicensis. In many respects species of Artibeus are obviously diverse, e.g., A. phaeotis has higher immunological affinities with other phyllosto- matid bats than with the related A. jamaicensis (Gerber and Leone, 1971) and moreover differs chromosomally from other species of Artibeus (Geno- ways and Baker, 1972). On the other hand, hemoglobins of all Artibeus species studied here are electrophoretically indistinguishable from each other and from other phyllostomatid bats (Valdivieso et al., 1969; Tamsitt and Valdivieso, 1969). Stenoderma rufum, one of the least known neotropical stenodermine bats (Jones et al., 1971), is similar to mainland South American phyllo- stomatids (Carollia, Artibeus, and Desmodus) but differs from other Puerto Rican phyllostomatid bats (A. jamaicensis, B. cavernarum, M. redmani, and FE. bombifrons) in albumin mobility. In number and mobilities of glob- ulin fractions, S$. rufum differs from all phyllostomatids studied and is more similar to the glossophagine bats M. redmani and G. soricina. Chro- mosomally, however, S. rufum is similar to Artibeus species (Baker and Lopez, 1970; Genoways and Baker, 1972) and moreover shares a species of listrophorid mite in common with Puerto Rican A. jamaicensis (Cruz et al., 1974). The electrophoretic pattern of hemoglobin of S. rufum is indistinguishable from Puerto Rican or mainland phyllostomatids (Valdi- vieso et al., 1969; Tamsitt and Valdivieso, 1969). Why the serum protein fractions of S. rufum are more similar to glossophagine bats, both insular and mainland, than to phylogenetically-related stenodermine bats remains to be determined. The genus Carollia, placed in the subfamily Carollinae of the Phyllo- stomatidae (Koopman and Jones, 1970), is similar to mainland Artibeus species, S. rufum, and D. rotundus in albumin mobility but differs from these taxa in number and mobilities of fractions in the globulin regions. Although electrophoretic properties of hemoglobins of C. perspicillata do not differ from other phyllostomatid bats (Tamsitt and Valdivieso, 1969), immunologically species of Glossophaga (Glossophaginae) and Carollia are more closely related than has been inferred from morphological evi- dence (Gerber and Leone, 1971). Consequently, the taxonomic status of the Glossophaginae and Carollinae warrants further study. Glossophaga soricina from South America and Monophyllus redmani from Puerto Rico (Glossophaginae) are similar in albumin mobilities but differ in number and mobilities of globulin fractions. Although similarities in chromosomes between species of Glossophaga and Phyllostomus were reported by Baker (1970), electropherograms of these taxa are strikingly different (Fig. 3). Fig. 3 Schematic diagram of patterns of serum proteins of neotropical bats (cellu- lose polyacetate). The pattern of normal human serum is given as a refer- ence. Fractions always present in the alpha and beta globulin regions are indicated by dark stippling; fractions that may be absent are represented by light stippling. O marks the origin; anode at left. Mobility of human albumin is indicated by a thin vertical line. Desmodus rotundus is similar to S. rufum, C. perspicillata, and all species of Artibeus examined except the insular A. jamaicensis in albumin mobility. The pattern of the serum electropherogram of D. rotundus is indistinguish- able from those individuals of A. lituratus that exhibit a complete set of five globulin fractions and all but indistinguishable from those A. cf. jamai- censis possessing the same number of globulin fractions. Although once considered to be a distinct family, Desmodontidae (Anderson and Jones, 1967), vampire bats are now considered to be a subfamily of the Phyllo- stomatidae (Koopman and Jones, 1970). Immunological results (Gerber and Leone, 1971), karyotypic information (Forman et al., 1968), and data from electrophoretic analyses of hemoglobin (Tamsitt and Valdivieso, 1969), as well as data presented here on serum proteins, support this taxonomic allocation. Of the molossids, insular Molossus fortis is similar to mainland M. molossus in albumin mobility, but the former differs from the latter by a larger number of fractions in the globulin regions. Although the molossid bats and the vespertilionid bat Eptesicus fuscus are similar to certain phyllostomatid bats in albumin mobility, these species differ from all phyllostomatid bats studied to date in electrophoretic properties of hemo- globin (Tamsitt and Valdivieso, 1969) and lactate dehydrogenases (Val- divieso et al., 1968). E. fuscus (Vespertilionidae) is similar to Molossus species in albumin mobility and in mobility of the two fractions in the beta region but differs by mobilities of the fractions in the alpha region. Data on serum protein concentrations (total protein) have been pub- lished on hedgehogs (Morris and Rudge, 1970), deer (Seal and Erickson, 1969), and other mammals, primarily domestic (Schalm, 1970). A wide range (6-11 g/100 ml) has been reported in total protein of mammals, and concentrations reported here (4.1-6.5 g/100 ml) for bats are toward the lower extreme of values (Table 1). Although differences among reports may result from analytical procedures, as most methods are influenced by variations in technique (King, 1964), reasons for reduced concentrations of protein in bat sera remain unexplained. The sera or plasma of many different mammals including rodents (Nadler, 1968; Dalby and Lillevik, 1969), artiodactyls (Nadler et al., 1967; Seal and Erickson, 1969), and cetaceans (Gallien et al., 1970) have been studied electrophoretically, and data have been applied to the systematics of groups at various levels of the taxonomic hierarchy. In some groups serum protein patterns are useful to distinguish species (van Tets and McT. Cowan, 1966; Petersen, 1968; and others), whereas in others they are phylogenetically conservative (Gallien et al., 1970). Certain frac- tions of sera have been demonstrated to be polymorphic within a species, particularly albumins, transferrins, haptoglobins, and other post-albumins (see Manwell and Baker, 1970). Few data, however, are available to compare the amount of protein variation in tropical and temperate species; moreover, except for immunologic analyses of some phyllostomatids (For- man et al., 1968; Gerber and Leone, 1971), no data exist on serum proteins of tropical bats except those presented here. 18 Bats are a suitable group for studies in biochemical systematics, for much of their taxonomy has been well defined by morphological characters. There are nonetheless many interesting taxonomic problems in neotropical Chiroptera, and the recent application of criteria from biochemistry, serology, and particularly karyology (see Baker, 1970) have helped eluci- date evolutionary relationships not revealed by conventional morphological characters. The Desmodontinae (vampire bats), once regarded as a dis- tinct family, is now recognized as a subfamily of the Phyllostomatidae (Koopman and Jones, 1970); evidence from karyotypes (Hsu and Ber- nirshke, 1967) and immunologic analyses (Forman et al., 1968) was instrumental in determining this affinity. Bats of the genus Sturnira, previously considered a separate subfamily (Sturnirinae) of the Phyllosto- matidae, were placed in the subfamily Stenoderminae (Koopman and Jones, 1970), an allocation supported by similarities of chromosomes (Gardiner and O'Neill, 1969), hemoglobins (Tamsitt and Valdivieso, 1969), and data presented here on serum proteins. Speculation or correlation between electrophoretic patterns and develop- mental processes has been intentionally avoided until the separated pro- teins can be more fully characterized. But the normal electropherograms of certain species of neotropical bats have been established, and knowledge of such electrophoretic patterns may be useful in other studies. Likewise, cellulose polyacetate patterns of sera of bats may have taxonomic value, for specics can be differentiated, and relationships between certain taxa are indicated. Here, for example, we pointed out potential problems con- cerning relationships among the Phyllostomatidae and the relationship of species of Phyllostomus to other phyllostomatids. Variations in patterns of individuals and populations were considerable but may provide information for intraspecific studies. A thorough analysis of quantitative differences within a single species is needed, as are immunological data to determine homologies of serum fractions among taxa. Summary Serum proteins of 75 bats of 18 species of the families Mormoopidae, Phyllostomatidae, Vespertilionidae, and Molossidae from Colombia, Puerto Rico, and Venezuela were compared by cellulose polyacetate electro- phoresis. Number of fractions varied from four to eight in electrophoretic patterns. Interspecific differences were observed in the total number of fractions and in their relative mobilities. Although varying in mobility among species, albumin, the most intense and rapidly migrating zone, and the fraction in the gamma globulin region were monomorphic in all species. Polymorphism occurred in the number of fractions in the alpha and beta regions of Phyllostomus hastatus, P. discolor, Glossophaga soricina, Arti- beus jamaicensis, A. cf. jamaicensis, A. lituratus, and Carollia perspicillata (Phyllostomatidae). Comparison of percentage composition of fractions of serum proteins by sex and age without specific allocations resulted in statistically insignifi- cant differences, except that the percentage of fractions in the alpha glob- 19 ulin region was significantly less in adult than in gravid females. Total protein content (g/100 ml) varied from 4.1-6.5, and differences by species or attributable to sex or age were negligible except that values for an adult Phyllostomus discolor were greater than those of a fetus. Based on similarities of albumin mobilities, four groups were evident: (A) Pteronotus parnellii (Mormoopidae); (B) the phyllostomatid bats Phyllostomus hastatus and P. discolor (Phyllostomatinae); (C) the phyllo- stomatid bats Glossophaga soricina and Monophyllus redmani (Glosso- phaginae), Sturnira lilium, Brachyphylla cavernarum, Artibeus jamaicensis (Stenoderminae), Erophylla bombifrons (Phyilonycterinae), the vesper- tilionid bat Eptesicus fuscus (Vespertilionidae), and the free-tailed bats Molossus molossus and M. fortis (Molossidae); and (D) the phyllosto- matid bats Artibeus cf. jamaicensis, A. lituratus, A. phaeotis, Stenoderma rufum (Stenoderminae), Carollia perspicillata (Carollinae), and Desmodus rotundus (Desmodontinae). Resumen Se comparan las proteinas de sueros de 75 ejemplares correspondientes a 18 especies de quirdépteros de las familias Mormoopidae, Phyllostoma- tidae, Vespertilionidae y Molossidae de Colombia, Puerto Rico y Vene- zuela por medio de electroforesis de poliacetato de celulosa. Cuatro a ocho fracciones son visibles en los electroferogramas de estos murciélagos. Simi- laridades y diferencias cuantitativas y cualitativas son bien aparentes. Diferencias interespecificas se observan en el numero total de fracciones, lo mismo que en sus mobilidades relativas. Albumina, la zona mas intensa y rapida, varia en ciertas de las especies estudiadas siendo en todos los casos monomorfica asi como lo es la fraccién correspondiente a la region de la gama globulina. Cuando se observa polimorfismo en las fracciones proteinicas del suero, éste occure Unicamente en el numero de bandas presentes en las regiones alfa y beta de Phyllostomus hastatus, P. discolor, Glossophaga soricina, Artibeus jamaicensis, A. cf. jamaicensis, A. lituratus y Carollia perspicillata (Phyllostomatidae). Comparaciones en el porcentaje de la composicion de las fracciones de estas proteinas por sexo o edad, sin tener en cuenta alocaciones especificas, demuestran solamente una diferencia estadistica significante: el porcentaje de fracciones en la regi6n alfa es inferior en hembras adultas que en aqué- llas gravidas. La concentraci6n total de proteinas (g/100 ml) varia de 4.1 a 6.5 sin observarse diferencias referentes a especies, sexo 0 edades excepto por los valores obtenidos en un Phyllostomus discolor adulto en el cual son mayores que aquéllos correspondientes a un feto de la misma especie. Basdndonos en similaridades de mobilidad de albiminas podemos dis- tinguir cuatro grupos diferentes: (A) Pteronotus parnellii (Mormoopidae) ; (B) los murciélagos filostomatidos Phyllostomus hastatus y P. discolor (Phyllostomatinae); (C) los filostomatidos Glossophaga soricina y Mono- phyllus redmani (Glossophaginae), Sturnira lilium, Brachyphylla caver- narum, Artibeus jamaicensis (Stenoderminae), Erophylla bombifrons 20 (Phylloncyterinae), el vespertiliénido Eptesicus fuscus (Vespertilionidae) y los molésidos Molossus molossus y M. fortis (Molossidae); y (D) los filostomatidos A. cf. jamaicensis, A. lituratus, A. phaeotis, Stenoderma rufum (Stenoderminae), Carollia perspicillata (Carollinae) y el vampiro Desmodus rotundus (Desmodontinac). Las propiedades electroforéticas de las proteinas del suero son taxo- némicamente importantes pero pueden representar un valor limitado como indicadores de relaciones filogenéticas en estos mamiferos. Acknowledgments We are grateful to Dr. Omar Linares for field assistance; to the personnel of the Instituto de Zoologia Tropical, Universidad Central, Caracas, for the use of laboratory facilities; to Dr. Gonzalo Medina Padilla, Director, Rancho Grande Biological Station, Venezuela, for his cooperation and for the use of facilities; and to the staff of the Department of Mammalogy, Royal Ontario Museum, for their assistance. We thank Dr. R. L. Peterson for aid in identification of specimens and for critically reading the manu- script. The illustrations were prepared by Mrs. Sophie Poray and Mr. Leighton R. Warren, Royal Ontario Museum. Our research was supported by the National Research Council of Canada, the Canadian National Sports- men’s Show, the Penrose Fund of the American Philosophical Society, the Ontario Department of University Affairs, and the Royal Ontario Museum. 21 Literature Cited ANDERSON, S. AND J. K. 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Gee r., ua , ie « 5 mse den ow waethe{ ©: ' 7 a an a>) ue Tk ' hese vere ee eee cat Vag 7 mit}, Pa ti 5 a el eel eli ee a a, i < er eee - mana = ISBN: 0-88854-153-8